Characterization of hydantoin-5-propionic acid amidohydrolase involved in ergothioneine utilization in Burkholderia sp. HME13
Characterization of hydantoin-5-propionic acid amidohydrolase involved in ergothioneine utilization in Burkholderia sp. HME13
复制标题
伯克霍尔德氏菌中参与麦角硫因利用的乙内酰脲-5-丙酸酰胺水解酶的表征。
DOI：
10.1093/bbb/zbad002
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发表时间：
2023
期刊：
影响因子：
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通讯作者：
Shin-ichiro Kato
中科院分区：
文献类型：
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作者：
Hisashi Muramatsu;Akihito Koujitani;Masaaki Yamada;Hiroki Maguchi;Takehiro Kashiwagi;Shin-ichiro Kato
在我们之前的研究中，ertABC基因编码麦角硫氨酸酶，硫代尿刊酸水合酶，3-（5-氧代-2-硫代咪唑烷-4-基）丙酸脱氢酶被确定，所有这些都可能参与麦角硫因利用伯克霍尔德氏菌。HME 13.在本研究中，我们确定了ertD基因编码的金属依赖性乙内酰脲-5-丙酸酰胺水解酶在该菌株。含Mn 2+的ErtD在45 °C和pH 8.5下显示出最大活性，并且在高达45 °C的温度下稳定。含Mn ~（2+）的ErtD对乙内酰脲-5-丙酸的Km和Vmax值分别为2.8mm和16 U/mg。实时荧光定量PCR（Real-time polymerase chain reaction，PCR）结果显示，在伯克霍尔德氏菌中，tD基因的表达水平较低。在麦角硫因培养基中培养的HME 13细胞比在Luria-Bertani（LB）培养基中培养的细胞高3.3倍。Burkholderiasp.在麦角硫因培养基中培养的HME 13细胞中，检测到0.018 U/mg，而在LB培养基中未检测到。因此，我们建议ErtD参与麦角硫因利用在这个菌株。
In our previous study,ertABCgenes encoding ergothionase, thiourocanate hydratase, and 3-(5-oxo-2-thioxoimidazolidin-4-yl) propionic acid desulfhydrase were identified, all of which may be involved in ergothioneine utilization ofBurkholderiasp. HME13. In this study, we identify theertDgene encoding metal-dependent hydantoin-5-propionic acid amidohydrolase in this strain. Mn2+-containing ErtD showed maximum activity at 45 °C and pH 8.5 and was stable at temperatures up to 45 °C. TheKmandVmaxvalues of Mn2+-containing ErtD for hydantoin-5-propionic acid were 2.8 mmand 16 U/mg, respectively. Real-time polymerase chain reaction (PCR) revealed thatertDexpression levels inBurkholderiasp. HME13 cells cultivated in ergothioneine medium were 3.3-fold higher than those in cells cultivated in Luria–Bertani (LB) medium. ErtD activity in the crude extract fromBurkholderiasp. HME13 cells cultured in ergothioneine medium was 0.018 U/mg, whereas that in LB medium was not detected. Accordingly, we suggest that ErtD is involved in ergothioneine utilization in this strain.